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Figure 5. Differential abundance analysis of the top 10 genera in mother and infant groups. Statistical significance was analyzed using
the Wilcoxon test. (A) Taxonomic differences of the top 10 genera by using relative abundance; (B) Taxonomic differences of top 10
*
genera by using absolute abundance. P < 0.05.
While our marine-derived spike-in sequences minimize overlap with gut microbes, this does not eliminate
the risk of differential amplification due to GC content, secondary structure, or mismatches at primer
binding sites. These are intrinsic to 16S amplification, not the spike-in itself. However, because all samples
receive identical spike-in DNA and undergo the same library preparation, the bias - though present - is
consistent across samples, allowing meaningful cross-group comparisons.
The natural variation in intestinal microbial loads is a significant and potentially clinically important feature
that is often overlooked in standard protocols. Technically, adding spike-in DNA directly to the sample
DNA can remove any potential errors introduced due to lysis of the spike-in bacteria, as might happen
